17alpha-chloro and 17alpha-bromo-progesterones



United States Patent O 3,087,938 17a-CHLOR0 AND 17oc-BROM0-PROGESTERONES Hans Reimann, Bloomfield, and David H. Gould, Leonia, N.J., assignors to Schering Corporation, Bloomfield, N.J., a corporation of New Jersey No Drawing. Filed Aug. 2, 1961, Ser. No. 128,681 6 Claims. (Cl. 260-3973) This invention relates to novel, therapeutically useful 3,20-diketopregnanes having an unsaturation in the A- ring, and to methods for their manufacture. In particular, this invention relates to 17a-substituted-9u,1lB-dihalogeno derivatives of A-ring unsaturated pregnane-3,20- diones which exhibit valuable progestational properties.

Novel pregnanes prepared by the process of our invention are the compounds represented by the following formula:

wherein X is a halogen having an atomic weight greater than 19; Y is a halogen having an atomic weight less than 126 and being at least as electronegative as X; Q is a member of the group consisting of hydrogen and methyl; and R is a member of the group consisting of methyl, halogen of atomic weight greater than 19 and less than 126, hydroxy and acyloxy. Our novel compounds are thus 9a,11fi-dihalogeno derivatives of 170:- substituted progesterones, Wot-substituted-l9-norprogesterones, Hot-substituted-l-allopregnene-3,ZO-diones and 17 u-substituted-1,4-pregnadiene-3 ,20-diones.

Illustrative of the 17a-acyl groups in the 17a-3CY1OXY derivatives contemplated by our invention are lower alkanoates such as formate, acetate, propionate, butyrate, isobutyrate, valera-te, isovalerate, pivalate, caproate, isocaproate, emanthate; substituted alkanoates such as fl-cyclopentylpropionate, cyclohexylacetate, ethoxyacetate, phenylacetate, phenoxyacetate; unsaturated acyl radicals such as acrylate and crotonate; aromatic acyl radicals such as benzoate and toluate, as well as the residues of dibasic acids such as succinate and phthalate and of fatty acids such as undecanoate and laurate. The term acyloxy thus includes acyl radicals of monocarboxylic and dicarboxylic acids containing up to 12 carbon atoms.

As stated heretofore, the llp-halogen present in a compound of the general formula or an analog thereof, must be at least as electronegative as the halogen present in the 9m-positi0n; fluorine being the most electronegative halogen, and iodine the least electronegative. Thus, a progesterone derivative of our invention containing a 3,087,938 Patented Apr. 30, 1963 9a-chloro group may possess an llfi-chloro or an 11,6- fluoro group but cannot contain an 11 fl-iodo or llfi-bromo group. This artificial restriction is imposed in view of the limitation of the manufacturing process.

Typical derivatives contemplated by our invention are progesterones such as 911,11[3-dichloro-l7a-hydroxyprogesterone and 9a-bromo-1LB-chloro-17m-hydroxyprogesterone and the 17-acetate and 17-caproate esters thereof, as well as other 17-substituted progesterones such as 9a,11fi,17actrichloroprogesterone, 9a-bromo-1 lfi-fluoro Hot-chloroprogesterone, 9a,17a-dichloro-1 1 13-fluoroprogesterone, L,- 1 1 p-dichloro-17a-bromoprogesterone, 9a-iOdO-1 lp-chlorol7a-methylprogesterone, 9ot,1 1 B-dichloro- 17a-methylpro gester one; and include their l-dehydro analogs and 19-norprogesterones such as 9a,1I/S-dichloro-l7a-hydroxy-19- norprogesterone 17-(fi-cyclopentylpropionate) and 9mbromo-l 1 tit-fillOIO-17oc-hYdIOXY 19 norprogesterone 17- acetate, and l-allopregnene derivatives exemplified by 9a,l lfl-dichloro-l7ot-hydroXy-1-alloprognene 3,20 dione 17-acetate and 9a-bromo-1lB-chloro-17 a-hydroxy-l-allopregnene-3,20-dione 17 -acetate. Although our compounds (with the exception of the 17-hydroxy substituted compounds which are valuable as intermediates) are, in general, valuable progestins, the 17a-acyloXy-dihalogenated progesterones of the general formula are the preferred species and, in particular, 9a,1lfl-dichloro-lh-hydroxyprogesterone 17-acetate.

Our novel compounds are prepared by reacting a 4,9 (11)-pregnadiene-3,20-dione substituted at the 17-carbon by hydroxy, acyloxy, halogen or methyl (or a similarly substituted 1,4,9 1 1 )-pregnatriene-3,20-dione or 1,9 1 1 allopregnadiene-3,20 dione) with a suitable halogenating agent. The starting compounds utilized in this invention are therefore exemplified by compounds such as 17a-hydroXy-4,9(ll)-pregnadiene 3,20 dione, 17a-acetoxy-4,9 l 1 pregnadiene-3,20-dione, l7ot-methyl-4,9 1 1 -pregnadiene-3,20-dione, 17oc-b10m0- 4,9(11) pregnadiene-3,20- dione, 17a-chloro-4,9(1l)-pregnadiene-3,20=-dione as well as the l-dehydro analogs of the foregoing, and pregnadicues such as 17a-hydroxy-4,9(11)-19-n0rpregnadiene 3,20-dione and 17a-hydroxy-l,9(1l)-allopregnadiene-3,- 20-dione 17-acetate.

The starting compounds thus necessarily possess a A bond and, in the case of 17 a-hydroXy-4,9( l 1)-pregnadiene- 3,20-dione (17a-hydroxy- 9(11) dehydroprogesterone), the process of preparing this 9(1l)dehydroprogesterone is described in the literature. The other aforementioned starting 9(1l)-dehydro starting compounds are prepared by a combination of processes analogous to those described in the literature. The 17a-acyloXy-9(11)-dehydroprogesterones (i.e. the 17a-acyloXy-4,9(11)-pregnadiene-3,20-diones) are prepared from 17a-hydroXy-9(11)- dehydroprogesterone according to known esterification techniques with a suitable acid such as acetic in the presence of trifiuoroacetic anhydride or with a suitable acid anhydride (such as acetic) in the presence of an acidic catalyst such as p-toluenesulfonic acid. When other acids, such as caproic or B-cyclopentylpropionic are used in place of acetic in these esterification procedures, the corresponding 17ec-6St61 is obtained, i.e. the 17ot-caproate and 17 a-fi-cyclopentylpropionate of t-hYdI0Xy 4,9(1l)- pregnadiene-3,20-dione.

Some 9(11)-dehydro starting compounds are prepared through the ll-hydroxylated progesterones, for example, 17a-methyl-9 l 1 -dehydroprogesterone 17u-methyl-4,9 (1l)-pregnadiene-3,ZO-dione) is prepared from Not-methyl-l16,21-dihydroxy-4-pregnene-3,ZO-dione 17u-methylcorticosterone) by an initial dehydration as efiected by reagents such as methanesulfonyl chloride in the presence of pyridine which yields, if methanesulfonyl chloride is the reagent used, l7a-methyl-21-hydroxy-4,9(l1)-pregnadiene-3,20-dione 2l-methanesulfonate. Conversion of the 21-ester to the Zl-desoxy compound is effected by treatment of the ester with, for example, sodium iodide in acetone yielding the corresponding 21-iodo derivative which, upon addition of acetic acid, for example, yields the requisite intermediate, 17tx-methyl-9(11)-dehydroprogesterone.

Another 17-substituted-9(l1)-dehydro starting compound prepared from a 2l-hydroxy intermediate is 170:- hydroxy 1,9(11)-bisdehydroprogesterone (17a-hydroxyl,4,9(11)-pregnatriene-3,20-dione). The starting compound 17a,21-dihydroxy-1,9( 11)-bisdehydroprogesterone 21-acetate (17a,21-dihydroxy-1,4,9(11)-pregnatriene-3,20- dione 21-acetate) is hydrolyzed to the corresponding 21-01, 170:,21 dihydroxy-1,9(11)-bisdehydroprogesterone (1704, 21 dihydroxy 1,4,9(11)-pregnatriene-3,ZO-dione). The latter is then treated with a sulfonating agent, such as ptoluenesulfonyl chloride in pyridine (or methanesulfonyl chloride in methylene chloride-pyridine) which yields the corresponding C-21-tosylate (p-toluenesulfonate) or mesylate (methanesulfonate). The tosylate is replaced by iodine by means of sodium iodide in acetone, the 21- iodide upon subsequent reaction with sodium bisulfite solution yielding the 21-desoxy compound l7a-hydroxy-1, 9(11) bisdehydroprogesterone (17a-hydroxy-1,4,9(11)- pregnatriene-3 ,20-dione) When an ll-hydroxy intermediate is not available, as in the case of 11 8-hydroxy-17a-bromoprogesterone, the hydroxyl group is introduced microbiologically into the ll-desoxy analog, 17a-bromoprogesterone, by means of a microorganism such as Curvularia lunata (N.R.R.L. 2380) by procedures analogous to those described in U.S. Patent No. 2,658,023. The llfi-hydroxy-lh-bromoprogesterone thus produced upon treatment with, for example, lithium bromide in glacial acetic acid yields 170cbromo 9(11)-dehydroprogesterone (l7oc-b10II10-4,9(11) pregnadiene-3,20-dione). In like manner, 17m-acetoxy 1-allopregnene-3,20-dione (17a-hydroxy-1-allopregnene-3, 20-dione 17-acetate) is ll-hydroxylated by means of Curvularia lmzata to give 11B,17a-dihydroxy-1-allopregnene-3,20diode 17-acetate which when dehydrated with an agent such as methanesulfonyl chloride in pyridine yields 17a-acetoxy-1,9( 11)-.allopregnadiene-3,20-dione.

Our 9(ll)-dehydro starting compounds may also be prepared by introducing an lla-hydroxyl group into an ll-desoxy progesterone through the action of a microorganism such as Rhizopus nigricans (A.T.C.C. 6227b) using procedures analogous to those described in U.S. Patent No. 2,602,769. Subsequent treatment of the llahydroxyprogesterone derivative so obtained with a sulfonyl chloride such as methanesulfonyl chloride or toluenesulfonyl chloride yields the corresponding lla-sulfonate which when treated with a base such as pyridine gives the desired 9(11)-dehydroprogesterone derivative.

The intermediate l7oc-ChlO1O-9 l 1)-dehydroprogesterone is obtained from 3fi-hydroxy-5-pregnene-20-one 3- acetate by employing known chemical techniques. Typically, 35-hydroxy-5-pregnene-20-one 3-acetate when treated wtih acetic anhydride and p-toluenesulfonic acid is enol-acetylated to give 3/3,20-dihydroxy-5,17(20)-pregnadiene-3,20-diacetate which, upon chlorination, by a standard procedure such as with chlorine in carbon tetrachloride yields 50:,65,17a-trichloro-3fl-hydroxypregnane- 20-one 3-acet-ate. Dehalogenation of the trichloro intermediate utilizing a reagent such as sodium iodide in acetone gives l7a-cl1loro-313-hydroxy-S-pregnene-ZO-one 3- acetate which is hydrolyzed and oxidized upon treatment with the organism Flavobaclerium dehydrogenans (Rutgers collection No. 130) in a manner analogous to the procedures outlined in Union of South African Patent No. 3462/55 to give l7a-chloroprogesterone. The double bond between C-9 and C-11 is then introduced into the 17a-chloroprogesterone through the corresponding 11- hydroxy-derivative which is prepared microbiologically, for example, by means of Rhizopus nigricans or C urvularia lunata. The 1lfl-hydroxy-l7a-chloroprogesterone thus obtained (i.e. Curvularia lzmam is the microorganism used) is dehydrated by procedures described heretofore to give the necessary intermediate, 17a-hydroxy-9(ll)- dchydroprogesterone 17 ot-hydroxy4,9 1 1 -pregnadiene- 3,20-dione).

The l-dehydro intermediates, i.e. 17oc-methyl-1,9( 11)- bisdehydroprogesterone l7a-methyl-l,4,9(11)-pregnatriene-3,20-dione) 17a-bromo-1,9(11)-bisdehydroprogesterone (17a-bromo-1,4,9(11)-pregnatriene-3,ZO-dione), 17achloro 1,9(11)-bisdehydroprogesterone (l7a-Chl01'O-l,4, 9(11) pregnatriene 3,30-dione), 17a-hydroxy-1,9(ll)- bisdehydroprogesterone (17a hydroxy-1,4,9(11)-pregnatriene-3,20-dione), and 17ot-acyloxy intermediates such as a acetoxy l,9(11)-bisdehydroprogesterone (l7a-hydroxy-1,4,9(11)-pregnatriene-3,20-dione 17-acetate are conveniently prepared from the corresponding 17a-substituted-9(11)-dehydroprogesterone by microbiological dehydrogenation with an organism such as, for example, Corynebacterium simplex (A.T.C.C. 6946) in a manner similar to that described in U.S. Patent No. 2,837,464, or by chemical oxidation through the use of such reagents as chloranil or selenium dioxide, or by halogenation followed by dehydrohalogenation.

Halogenating agents suitable for use in our process are:

(1) Molecular halogens having a molecular weight greater than 38 and less than 253. This class includes heteroatomic halogen molecules such as iodine monochloride as well as isoatomic halogen molecules such as chlorine and bromine. The molecular halogen employed may be used alone or in admixture with a halide anion. In reactions employing isoatomic molecular halogens, an anion corresponding to the molecular halogen is used. For example, in reactions using molecular chlorine, a suitable chloride salt such as lithium chloride or hydrochloric acid are suitable anion sources. In reactions which employ a heteroatomic molecular halogen, a source of the more electronegative anion is used, i.e. a suitable chloride salt or hydrochloric acid is used in a reaction employing iodine monochloride.

(2) Addition compounds of molecular halogens, said molecular halogens having a molecular weight greater than 38 and less than 253. Examples of such addition compounds are pyridinium bromide perbromide, pyridinium chloride perchloride, dioxane dibromide, iodobenzene dichloride, and the like.

(3) N-haloamides in admixture with a halide anion, the halogen in the halide being at least as electronegative as the halogen cation in the N-haloamide, and the combined molecular weight of said halogens being greater than 38 and less than 253. N-haloamides used in these mixtures are such as N-chlorosuccinimide, N-bromoacetamide, dimethyl-N,N-dibromohydantoin, N-iodosuccinimide and the like. Examples of mixtures of N- haloamide and a halide are such as N-chlorosuccinimide and hydrogen chloride; N-bromoacetamide and hydrogen bromide; N-bromoacetamide and lithium chloride; N- bromoacetamide and hydrogen fluoride; N-iodosuccinimide and sodium chloride. These mixtures may have more than one source of halide ion such as in the reagent combination N-chlorosuccinimide, hydrogen chloride and lithium chloride.

Molecular halogens (both isoatomic and heteroatomic) such as chlorine, bromine and iodine monochloride, and molecular halogen addition compounds such as iodobenzene dichloride or pyridinium bromide perbromide employed in our process may be used directly in their commercially available form, in which case they are added directly to the reaction mixture either alone or in a suitable non-reacting solvent such as acetic acid or tetrahydrofuran. The halogenating agent is preferably used in approximately equivalent quantities to that of the steroid.

Alternatively, the halogenating reactant may be prepared in situ. For example, 0.9-1.2 equivalents of halogen cation (based on the amount of steroid) such as obtained from N-bromosuccinimide is added to a reaction mixture containing a halogen anion of greater or equal electronegativity than the cationic reagent. The halogen anion may be supplied by a hydrohalic acid or by a salt such as sodium bromide, lithium chloride, potassium fluoride, or by mixtures of a hydrohalic acid with an alkali metal halide. Whenever an acid is the source of a halogen anion, approximate theoretical quantities are generally used; whereas if a salt is used as the halogen anion source it may be present in large excess. Such an excess of anion is frequenly useful to obtain an increased yield or greater purity of product.

Thus, our novel compounds may be prepared by any one of several methods. For example, 9a,11fldichlorol7a-hydroxyprogesterone l7-acetate may be prepared from 17a-hydroxy 9(11) dehydroprogesterone 17-acetate by utilizing such reagents and combinations as (1) chlorine and lithium chloride, (2) N-chlorosuccinimide and lithium chloride, (3) sodium chloride, N-chlorosuccinimide together with hydrogen chloride, (4) chlorine alone, (5) chlorine and hydrogen chloride, and (6) iodobenzene dichloride.

An alternate route for preparing the 17a-acyloxy compounds of our invention is halogenating 17a-hydroxy- 9(ll)-dehydroprogesterone (or the l-dehydro analog or 19-nor analog thereof or a 17ot-hydroxy1,9(l1)-allopregnadiene-3,20-dione) by any of the above described halogenating methods of our invention, with subsequent esterification of the '9a,1lfi-dihalogenod7a-hydroxyprogesterone (or the l-dehydro or 19-nor analog or the 90:, 11,8 dihalogeno-17a-hydroxy-l-allopregnene-3,ZO-dione) thereby obtained. In this manner l7a-hydroxy-9(11)-dehydroprogesterone, upon chlorination with a reagent such as chlorine in carbon tetrachloride in the presence of pyridine, yields 9a,1IB-dichloro-l7a-hydroxyprogesterone which, when esterified with acetic acid and trifluoroacetic anhydride, for example, gives 9u,llB-dichl-or0-l7u-hydroxyprogesterone 17-acetate. Other acids may be substituted for acetic acid in the esterification step to obtain.

the corresponding l7a-acyloxy dihalogenated progesterone.

Our novel halogenating process is also useful in preparing the 1,4-pregnadienes of our invention. Thus, a 17- substituted-90 1 1p-dihalogeno-l-dehydroprogesterone such as, for example, 9a,llfi-dichloro-17a-methyl-1-dehydroprogesterone is prepared from l'7a-methyl-1,9(11)-bisdehydroprogesterone by utilizing any one of the six reagents and combinations listed previously. In addition, our novel halogenated l-dehydroprogesterones are obtained from the corresponding halogenated progesterones by microbiological or chemical dehydrogenation techniques as heretofore described. Thus, 9a,1lfl-dichloro-lh-methylprogesterone is converted to 9a,1lfl-dichloro-17a-methyll-dehydroprogesterone by the action of a culture of the microorganism Corynebactei'ium simplex.

Our process, whereby a 17-substituted-9(l1)-dehydroprogesterone is converted to a 17u-substituted-9u,1lfi-dihalogeno derivative is generally carried out in the presence of a non-reacting solvent at temperatures ranging from 5 C. to 50 C. with reaction times varying from one-half to 24 hours, depending on the reagents involved. It is preferred that the halogen cation source be present in amounts of 0.9-1.2 equivalents based upon the amount of steroid. The choice of solvent in each reaction is naturally determined by the solubility of the reactants in the process. A preferred reaction condition employs glacial acetic acid as the sole or major solvent with the reaction being carried out at room temperature for a period of approximately two hours.

Although glacial acetic acid is frequently the preferred solvent, other solvents are used in our dihalogenation process either alone or in combination with acetic acid. Other solvents which may be employed include lower aliphatic acids such as propionic and diethylacetic, halo- 6 gen'ated hydrocarbons such as methylene chloride and chloroform, saturated ethers such as tetrahydrofuran and dioxane, and inert solvents such as dimethylsulfoxide, as well as suitable mixtures of these solvents.

Further, when carrying out the above-described process in a halogenated hydrocarbon solvent such as methylene chloride, chloroform or carbon teterachloride, the yield and purity of the dihalogenated progesterone obtained is greatly increased by adding pyridine to the reaction mixture. Preferably from one to five equivalents of pyridine is added, based on the amount of steroid being converted. Other organic basic agents which may also be used in conjunction with a halogenated hydrocarbon solvent are aromatic tertiary amines such as lutidine, collidine, alkyl substituted pyridines, and the like. The reaction is usually carried out intitially at -20 C. with subsequent warning to about room temperature. The reaction time may be as short as five minutes or as long as several hours. In general the optimum reaction time is about one hour.

Our process whereby a l7-substituted-9(11)-dehydroprogesterone is converted to a l7-substituted-9a,11B-dihalogeno progesterone is also applicable, as mentioned previously, to 17-substituted-9(11)-dehydro-19-norprogesterones. With the aforementioned 9(11)-dehydro-19- norprogesterones there are produced the novel progesta tional agents, 9oc-X-1lfl-Y-l7oc-R-19-11OIPIOgESteIOI1fiS wherein X, Y, and R are as heretofore described. Thus, 17a methyl 9(11) dehydro-l9-norprogesterone (17ccmethyl 19 nor 4,9(11)-pregnadiene-3,20-dione), cbromo 9(11)-dehydro-19-norprogesterone (l7oc-br0m0- 19 nor 4,9(11)-pregnadiene-3,20-dione), 17a-chloro-9 1 1 -dehydro-19-norprogesterone l7a-chloro-19-nor-4, 9 1 1 pregnadiene-3,20-dione) l7ot-hydroxy-9 11 )-dehydro 19 norprogesterone (17a-hydroxy-19-nor-4,9(11)- pregnadiene-3,20-dione) and 17ot-acyloxy-9(11)-dehydro- 19-norprogesterones 17 a-acyloxy-19-nor-4,9 11)-pregnadiene-3,20-diones), when reacted with chlorine in carbon tetrachloride in the presence of pyridine, for example, are converted to their respective 9a-11B-dichloro derivatives.

When preparing a 9a,11j3-dihalogenated l7u-acyloxy- 19-norprogesterone by our process the halogenation step is preferably performed on the corresponding l7a-hydroxy- 9(l1)-dehydro intermediates prior to esterification of the 17-hydroxy group. Thus, 17a-hydroxy-9(11)-dehydroprogesterone is first converted to a 9oc,1l,6-dihalogenated- 17a-hydroxy-19-norprogesterone which is then esterified by methods heretofore described to give the 9(11)-dihalogenated-l7a-acyloxyprogester0ne.

Our novel dihalogenated 17-substituted progesterones, their l-dehydro and l9-nor analogs, as well as the dihalogenated '17-substituted-l-allopregnenes of our invention (except those having a hydroxy group at C-17) are very active progestational agents being specific in their action, and devoid of androgenic, estrogenic or corticoid activity. That our compounds have progestational activity at all is surprising in view of the well known fact that substitution in ring-C of progesterone and its derivatives such as, for example, ll-hydroxyprogesterone and the ll-acyloxy derivatives thereof eliminates progestational activity or reduces it to such an extent that the compounds are useless for therapeutic purposes. Our ring-C halogenated-17-substituted progesterones, on the other hand, are significantly more active than progesterone by the intramuscular route. Moreover, orally our compounds are several times as active as ethisterone, the known standard progestational agent.

In addition to being active both orally and intramuscularly, our halogenated progestins possess the added advantage of having a minimum effect on water, sodium and potassium metabolism. Thus, they may be administered without causing the usual side efiects associated with electrolyte imbalance.

Our therapeutically active compounds are useful for the treatment of conditions requiring progestational agents such as the maintenance of pregnancy, or treatment of functional dysmenorrhea, premenstrual tension, habitual or threatened abortion. When administered orally, our compounds are preferably used in the form of tablets containing from 1 to 10 mg. together with the excipients such as starch or milk sugar. For subcutaneous and intramuscular administration, solutions or suspensions of our compounds with a non-toxic liquid vehicle are used. The dosage required may vary with the indications being treated and may range from about 5 to 25 mg. daily.

Our halogenated progestins are also valuable in the veterinary field for treating conditions in both large and small animals which require a progestational agent. In breeding animals, for example, our compounds are useful in preventing threatened abortion. Additional uses are in controlling egg and milk production by regulating the cycle of chickens and cows by the administration of our progestational agents.

The 17a-hydroxy compounds of our invention, i.e. the l7a-hydroxy 90:,1119 dihalogenoprogesterones, the l7a-hydroxy-9a,llfi-dihalogeno-l dehydroprogesterones, the 17a-hydroxy-9a,1IB-dihalogeno-l9-norprogesterone, and the '17a-hydroxy-9a,1lfi-dihalogeno-l-allopregnene- 3,20-dior1es, are valuable mainly as intermediates in the preparation of the corresponding 17a-acyloxy compounds, which are active progestational egents.

The following examples are illustrative of the procedures employed in preparing the compounds of this invention, but are not to be construed as limiting the scope thereof; the scope of our invention being limited only by the appended claims.

This application is a continuation-in-part of our copending application, Serial No. 113,089, filed May 29, 1961.

EXAMPLE 1 1 7a-Hydr0xy-4,9(1 1 -Pregnadiene-3,20-Dine 1 7-Acetate To a solution of 5.0 g. of 17a-hydroxy-4,9(11)-pregnadiene-3,20-dione in 50 ml. of acetic acid there is added 10 ml. of trifluoroacetic anhydride. The solution is heated on the steam bath for 45 minutes, then is poured into ice-water. A solid separates which is filtered and crystallized from acetone-ether to give 17a-hydroxy- 4,9(11)-pregnadiene-3,20-dione 17-acetate, M.P. 238- 241 C., [u] +O (chloroform);

Analysis.-Calcd. for C H O C, 74.56; H, 8.16. Found: C, 74.64; H, 8.07.

EXAMPLE 2 l 7a-Hydr0xy-4,9 (11 )-Pregnadiene-3,20-Dione 1 7 -Capr0ate A mixture of 1.0 g. of 17a-hydroxy-4,9(11)-pregnadiene3,20-dione and 5 ml. of caproic acid is heated at 80 C., in the presence of 1.0 ml. of trifluoroacetic anhydride, for 45 minutes. The mixture is poured into icewa ter and the excess acid neutralized with sodium carbonate. The crude reaction mixture is extracted with methylene chloride. The organic extracts are combined and concentrated to a residue which is chromatographed on Florisil. The material eluted with -25% ether in hexane is combined and crystallized from pentane to yield 17a-hydroxy-4,9(11)-pregnadiene-3,20-dione 17- caproate, M.P. 126-128 C.

X2552}! 239 my (e=l6,800), [u]' +42 (chloroform).

Analysis.Calcd. for C H O C, 76.02; H, 8.98. Found: C, 76.11; H, 8.94.

8 EXAMPLE 3 1 7a-Hydr0xy-4,9(1l -Pregnadiene-3,20-Dione 1 7 B-Cycl0pentylpr0pi0nate) To a solution of 500 mg. of 17a-hydroxy-4,9(11)- pregnadiene-3,20-dione in 10 ml. of fi-cyclopentylpropionic acid and 2.5 ml. of 18-cyclopentylpropionic anhydride in a vessel flushed with nitrogen, there is added 200 mg. of p-toluenesulfonic acid with stirring at a temperature of about 20 C. The reaction mixture is allowed to stand for 4 hours, then poured into ice-water. Excess acid is neutralized with sodium carbonate, and the crude mixture is extracted with ether. The ether extracts are combined, washed with water, dried over magnesium sulfate, filtered, and concentrated to a residue. The residue is crystallized from methanol to give 17ot-hydroxy-4,9(1l)- pregnadiene-3,20-dione l7-(fi-cyclopentylpropionate) max.

EXAMPLE 4 9a,]1fl-Dichl0ro-l 7a-Hydroxyprogesterone 1 7 -A cetate A solution of l g. of l7a-hydroxy-4,9(ll)-pregnadiene- 3,20-dione 17-acetate (the compound of Example 1) and 4 g. of lithium chloride in. 40 ml. of glacial acetic acid is cooled to about 10 C. and there is added 250 mg. of hydrogen chloride in 1 ml. of tetrahydrofuran followed by 395 mg. of. 96% N-chlorosuccinimide. The solution is stirred at room temperature for 20 minutes in the absence of light, then is poured into ice-water with stirring. A precipitate forms which is filtered, washed with water, triturated with ether and crystallized twice from acetone-hexane to give 901,11/3-dichloro-17a-hydroxyprogesterone 17-acetate, M.P. 224-233 C. dec., [ab-l- 129 (chloroform) AMeOH m... 238 III/L e=17,000.

A'nalysis.-Calcd. for C H O Cl C, 62.58; H, 6.85; Cl. 16.06. Found: C. 62.63; H, 7.03; CI, 15.90.

Alternatively, the compound of this example is prepared by adding a solution of 200mg. of chlorine in 30 m1. of acetic acid to a mixture of 1 g. of 17a-hydroxy- 4,9(11)-pregnadiene-3,20-dione 17-acetate and 4 g. of lithium chloride in 20 ml. of glacial acetic acid. The mixture is stirred at room temperature for 3 hours, then poured into ice-water with stirring. A solid product precipitates which is filtered, washed with water, triturated with ether, and crystallized from acetone-hexane to give 9a,l lfi-dichloro-17a-hydroxyprogesterone 17-acetate.

A second alternative for the preparation of the compound of this example isby reacting 1 g. of 17a-hydroxy- 4,9(11)-pregnadiene-3,20-dione l7-acetate dissolved in 30ml. of carbon tetrachloride with 1.8 m1. of chlorine in carbon tetrachloride (11'1 mg./ml.) in the presence of 0.65 ml. of pyridine at -20 C. The mixture is stirred at -20 C. for 15 minutes, then is allowed to warm to room temperature over a period of one-half hour. The solution is filtered and the filtrate concentrated in vacuo to an oily residue. Trituration of this residue with ether yields solid material which is crystallized from acetonehexane to give 9a,1lfl-dichloro-17a-hydroxyprogesterone 17-acetate.

EXAMPLE 5 9a-Br0m0-Jlfl-Chlor0-1 7a-Hydr0xyprogester0ne 1 7-A cetate One gram of 17a-hydroxy-4,9(1l)-pregnadiene-3,20- dione l7-acetate (the compound of Example 1) and 4 g. of lithium chloride is dissolved in 50 ml. of glacial acetic acid and there is added 420 mg. of N-bromoacetamide. The mixture is stirred at room temperature and a slow stream of gaseous hydrogen chloride is passed over the surface until the solution begins to darken (10-30 seconds). The hydrogen chloride gas is removed and the solution is stirred in the dark at room temperature for about 10 minutes. The solution is poured into ice-water with stirring and the resultant solid is filtered, washed with water, and crystallized from acetone-hexane to give 9a-bromo-llfl-chloro-l7u-hydroxyprogesterone 17 acetate, M.P. 124-129 C. dec., [ad -142 (chloroform) Analysis.--Calcd. for C H O B1'C1: C, 56.85; H, 6.22; Br, 16.45; Cl, 7.30. Found: C, 56.83; H, 6.28; Br, 16.07; Cl, 7.35.

EXAMPLE 6 QOL-BI'OITlO-J 1 ,B-Fluoro-I 7a-Hydr0xyprogesterone 1 7-A cetate To a solution of 1 g. of 17a-hydroxy-4,9:(11)-pregnadiene-3,20-dione l7-acetate (the compound of Example 1) and 425 mg. of N-bromoacetamide in 50 ml. of diethylacetic acid there is added a solution of 500 mg. of hydrogen fluoride in 4.7 ml. of chloroform-tetrahydrofuran. The solution is stirred at room temperature for 2 hours, then poured into ice-water with stirring. The acid is neutralized by addition of sodium bicarbonate and a resinous precipitate forms from which the aqueous solution is decanted. The precipitate is then dissolved in methylene chloride and the solution is washed with 5% aqueous sodium hydroxide, then with water, is dried over magnesium sulfate, filtered and concentrated in vacuo. The resultant residue is triturated with pentane, filtered and crystallized from methylene chloride-pentane to give 9a-bromo-l1p-fluoro-l7a-hydroxypr-ogesterone l7-acetate, MJP. 176-180 C. dec.,

Analysis.Calcd. for C H O BrF: Br, 17.03; F, 4.05. Found: Br, 17.56; F, 3:85.

EXAMPLE 7 90:, 11 fl-Di ch 1 0r0-1 7 a-H y droxy progesterone 1 7-Capr0ate 17a-hydroxy-4,9 l 1 -pregnadiene-3,20-dione 17 -caproate (the compound of Example 2) is reacted with 165 mg. of N-chlorosuccinimide, hydrogen chloride and lithium chloride in the manner described in Example 4. The resultant product is isolated and purified in the described manner to give 9a,1lfi-dichloro-l7a-hydroxyprogesterone 17-caproate,

EXAMPLE 8 9a,] 1 fl-Dichloro-l 7 a-H ydroxy progesterone 1 7-( fl-Cyclopentylpropionate) 17 a-hydroxy-4,9( l 1 )-pregnadiene 3,20 dione 17-(5- cyclopentylpropion-ate), the compound of Example 3, is reacted with chlorine and lithium chloride in the manner of Example 4 and the resultant product isolated and purified to give 9a,1LB-dichloro-17a-hydroxyprogesterone 17-(fi-cyclopentylpropionate),

KA SE 238 my.

EXAMPLE 9 9a,]1,B-Dibrom0-1.7a-Hydr0xyprogesterone 1 7-A cetate To a solution of 1 g. of 17a-hydroxy-4,9'(11)-pregnadiene-3,20-dione 17-acetate (the compound of Example 1) and 4 g. of lithium bromide in 50 ml. of acetic acid there is added 400 mg. of N-bromoacetamide followed by a solution of 220 mg. of hydrogen bromide in 4.3 ml. of acetic acid. The mixture is stirred at room temperature for one hour, then is poured into ice-water. A solid forms which is filtered, washed with water, then crystallized and recrystallized twice from acetone-hexane to give ,1lp-dibromo-17u-hydroxyprogesterone 17* acetate,

AM? 241 my EXAMPLE 1o 9a-I0do-1 1 fi-Chloro-I 7 a-H ydroxy progesterone 1 7-A (:etate To a solution of l g. of 17a-hydroxy-4,9(11)-pregchloride, the solution is warmed, decolorizing carbon added, then filtered. Pentane is added to the clarified methylene chloride solution and a precipitate is formed which is filtered and crystallized from acetone-hexane to give 9a-iodo-1lfi-chloro-l7a-hydroxyprogesterone l7- acetate.

EXAMPLE 11 9a-Chl0r0-1 1 fl-Fluoro-I 7 -H y droxy progesterone 1 7-Acetate To a solution of 500 mg. of 17a-hydroxy-4,9(l1)-preg nadiene-3,20-dione l7-acetate (the compound of Example l) in 25 ml. of diethylacetic acid there is added 195 mg. of N-chlorosuccinimide followed by a solution of '620 mg. of hydrogen fluoride in 3.2 m1. of a mixture of tetrahydrofuran and chloroform. The reaction mixture is stirred at room temperature for 48 hours, then is poured into an aqueous sodium carbonate solution. The mixture is extracted with methylene chloride. The organic extracts are combined and evaporated to a residue which is chromatographed on silica gel. The product eluted with 20% ether in hexane is crystallized with acetone-hexane to give 9a-chloro-11fl-fluoro-17-hydroxyprogesterone 17-acetate,

238 m, EXAMPLE 12 9a-l0d0-l1/3-Fluoro-1 7-Hydr0xypr0gester0ne .1 7-Acetate To a solution of 500 mg. of 17a-hydroXy-4,9(11)-pregnadiene-3,20-dione 17-acetate (the compound of Example 1) and 3 .g. of potassium fluoride in 40 ml. of dimethylsulfoxide there is added 320 mg. of N-iodosuccinimide. The reaction mixture is stirred at room temperature for 16 hours, poured into ice-water and extracted with methylene chloride. The combined organic extracts are warmed, decolorizing carbon added, then filtered, and the filtrate concentrated in vacuo. The resultant residue is chromatographed on silica gel. The fraction eluted with 50% ether-hexane yields 9a-iodo-llfl-fluoro- 17-hydroxyprogesterone l7-acetate.

EXAMPLE 13 17a-Hydroxy-1,4,9 (11 -Pregnatriene-3,20-Dione (A) 17a,21-DIHYDROXY -1,4,9(lb-PREGNATRIENE- a o-moms A solution of 2 g. of 17a,21-dihydroxy-l,4,9(11)-pregnatriene-3,20-dione ZI-acetate in ml. of a 3:1 methanol chloroform mixture is chilled in ice to which there is added dropwise 52 ml. of 0.1 N sodium hydroxide solution (one equivalent). The reaction mixture is stirred at 0 C. for 10 minutes, then diluted with water and extracted with methylene chloride. Evaporation of the solvent and crystallization of the residue from ace- 11 tone-hexane affords l7a,2l-dihydroxy-l,4,9(ll)-pregnatriene-3,20-dione, M.P. 220-228C. (dec.);

Analysis.Calcd. for C H O C, 73.66; H, 7.66. Found: C, 73.58; H, 7.54.

(B) 17a-HYDROXY-1,4,9 (11)-PREGNATRIENE 3,20-DIONE A solution of 10 g. of 17a,21-dihydroxy-1,4,9(11)- pregnatriene-3,-20-dione (the compound of Example 13A) in 100 ml. of dry pyridine is chilled in a Dry Ice-acetone bath and there is added a solution of 6 g. of p-toluenesulfonyl chloride in 50 ml. of methylene chloride. The mixture is stirred in the cold bath for 2 hours and then kept at -20 C. for 24 hours. At the end of this time, the reaction mixture is diluted with methylene chloride and the solution washed first with water, then with 10% sulfuric acid, 10% sodium bicarbonate, very dilute hydrochloric acid and finally with water. The solution is dried over a magnesium sulfate, filtered and concentrated to a residue which is dissolved in acetone. The acetone solution is warmed on the steam bath, decolorizing carbon added, and the solution filtered. To the clarified soluiton there is added a warm solution of 10 g. of sodium iodide in acetone. After :heating this mixture on the steam bath for five minutes, three ml, of acetic acid is added, and the mixture is warmed briefly, and then aqueous sodium bisulfite. The resulting pale yellow solution is poured into Water and the. resulting precipitate is filtered and washed with water, and crystallized from acetone-ether to give 17u-hydroxy-1,4,9 (11)-preguatriene-3,20dione, M.P. 233-235 C.

3522 238 u, e=15,600 [a] 19 (chloroform).

EXAMPLE 14 1 7a-Hydr0xy-1,4,9(1'1)-Pregnatriene-3,20-Dione 1 7-Acetate A solution of 5 g. of 17a-hydroxy-1,4,9(I11)-pregnatriene-3,2O-dione (the compound of Example 13) is dissolved in 50 ml. of glacial acetic acid and of trifluoroacetic anhydride. The solution is left at room temperature for 24 hours, then poured into ice-water. A solid separates which is filtered, washed with water, and crystallized from acetone-hexane to give 1'7a-hydroxy1,4,9 l 1 )-pregnatriene-3',20-dione l7-acetate,

AMeOH 23 mp max.

EXAMPLE 9a,11fi-Diclzl0r0-1 7u-Hydr0xy-1,4-Pregnadiene- 3,20-Dione 17-Acetate One gram of l7a-hydroxy-1,4,9(11)-pregnatriene-3,20- dione 17-acetate (the compound of Example 14) is reacted with 395 mg. of N-chl'orosuccinimide in the presence of hydrogen chloride and lithium chloride in the manner of Example 4 to give 9a,1 1 3-di( hl0lO-17u-hydroxy-l,4-pregnadiene-3,'2.0dione 17-acetate,

mos 7 m max.

The flask is incubated at 28 C. for 24 hours. A second 300 ml. Erlenmeyer flask containing mg. of sterile 17a hydroxy 1,4,9(ll) pregnatriene 3,20- dione 17-acetate in 5.0 ml. acetone is inoculated with the 24-hour culture of Corynebacterium simplex (A.T.C.C. 6946). The culture-containing steroid solution is incubated for 48 hours at 28-30" C.

After termination of the transformation period, the pH is 7.2-7.3. The culture is now directly extracted with 3 equal volumes of CHCl the solvent volumes combined and concentrated on a steam bath to a residue which is crystallized from acetone-hexane to give 9a,11f3- dichloro-l7a-hydroxy-1,4-pregnadiene-3,20-dione l7-acetate.

A second alternative method for the preparation of the compound of this example is as follows.

17a-hydroxy-1,4,9( ll)-pregnatriene 3,20 dione (the compound of Example 13B) is reacted with 220 mg. of chlorine gas in carbon tetrachloride in the presence of pyridine in the manner of the alternative procedure of Example 4 to give 9a,1lfl-dichloro-17a-hydroxy-1,4- pregnadiene-3,20-dione, M.P. 235-239 C. dec.

AMQOH 238 mu EXAMPLE 16 9a-I0do-1lfl-Chloro-17u-Hydr0xy-1,4-Pregnadiene- 3,20-Dione 1 7-A cetate To a solution of 1.0 g. of l7a-hydroxy-1,4,9(1l)- pregnatriene-3,20-dione 17-acetate (the compound of Example 14) in 40 ml. of tetrahydrofuran is chilled to 0 C., there is added 3 drops of perchloric acid and 3. g. of lithium chloride, followed by dropwise addition of a solution of 470 mg. of iodine monochloride in 5 ml. of tetrahydrofuran. The reaction mixture is stirred at room temperature for 5 hours, then poured into icewater. A gummy precipitates forms which is dissolved in methylene chloride. The solution is warmed, decolorizing carbon added, then filtered. Pentane is added to the clarified methylene chloride solution and a solid forms which is filtered and crystallized from acetonehexane to give 9a-iodo-11/8-chloro-17a-hydroxy-l,4-pregnadiene-3,20-dione 17-acetate.

Alternatively, 9a-iodo-1 l B-chlorol 7a-hydroxyprogesterone 17-acetate (the compound of Example 10) is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give 9a-iOdO-l1B-Chl01O-l7ahydroxy-l,4-pregnadiene-3,20-dione 17-acetate.

EXAMPLE l7 9a-Chl0r0-J Ifi-Fluoro-I 7a-Hydr0xy-1,4-Pregnadiene- 3,20-Di0ne 1 7 -A cetate 17oz hydroxy 1,4,9(11) pregnatriene 3,20 dione 17-acetate (the compound of Example 14) is reacted with N-chlorosuccinimide and hydrogen fluoride, and the resultant product isolated and purified in the manner described in Example 11 to give 9a-chloro-l1fi-fluoro- 17a-hydroxy-1,4-pregnadiene-3,20dione l7-acetate.

Alternatively, 9a-chloro-1 1fi-fluoro-17u-hydroxyprogesterone 17-acetate (the compound of Example 11) is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give 9oc-ChlO1'0-11fi-flUOIO-l7ahydroxy-l,4-pregnadiene-3,20-dione 17-acetate.

13 EXAMPLE 1:;

9a-Brom0-1 1fi-Chl0ro-17a-Hydroxy-1,4-Pregnadiene- 3,20-Dione 17-Acetate 17u-hydroxy-1,4,9(11)-pregnatriene 3,20 dione 17- acetate (the compound of Example 14) is reacted with N bromoacetamide, lithium chloride and gaseous hydrogen chloride in acetic acid in the manner described in Example 5 to give 9u-brom0-LIB-chloro-lh-hydroxy-l,4- pregnadiene3,20-dione 17-acetate.

Alternatively, 9a-br0m0-1 1fl-'Ch1OIO-17a hydroxyprogesterone 17-acetate (the compound of Example 5) is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give 90t-bI0lI10-11fi-Ch1OIO-170thydroxy-1,4-pregnadiene-3,20-dione 17-acetate.

EXAMPLE 19 9a-Br0m0-l1p-Fluoro-I7a-Hydr0xy-1,4-Pregnadiene- 3,20-Di0ne 1 7-Aceiate 17a-hydr0xy-1,4,9( 11 -pnegnatriene-3,20-dione 17-acetate (the compound of Example 14) is reacted with N-bromoacetamide in the presence of hydrogen fluoride and the resultant product isolated and purified in the manner described in Example 6 to give 9a-bromo-11fifluoro-17a-hydroxy-1,4-pregnadiene-3,20-dione 17 acetate.

Alternatively, 9a-bromo-11/i-fluoro-17a hydroxyprogesterone 17-acetate (the compound of Example 6) is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give 9a-bromo-1 1,3-fluoro-l7ahydroxy-l,4-pregnadiene-3,20 dione 17-acetate.

A second alternative procedure is as follows.

17a-hydroxy-1,4,9(11)-pregnatriene 3,20 dione (500 mg), the compound of Example 13, is reacted with 210 mg. of N-bromoacetamide in the presence of hydrogen fluoride in diethylacetic acid in the manner of Example 6. The product is isolated and purified in the described manner to give 9a-bromo-11fi-fluoro-17a-hydroxyprogesterone, which is then esterified with acetic acid in the presence of trifluoroacetic acid in the manner described in Example 14 to give 9a-bromo-1lB-fiuoro-lh-hydroxy-l,4- pregnadiene-3,20-dione 17-acetate.

EXAMPLE 20 9a,11;8-Dibr0m0-1 7oc-Hydr0xy-1,4-Pregnadiene- 3,20-Di0ne 17-Acetate 17a-hydroxy 1,4,9(11) pregnatriene-3,20-dione l7- acetate (the compound of Example 14) is reacted With N- bromoacetamide, lithium bromide and hydrogen bromide in acetic acid and the resultant product isolated and purified in the manner described in Example 9 to give 9'a,11,8- dibromo-17a-hydroxy-1,4-pregnadiene-3,ZO-dione l7-acetate.

Alternatively, 90,11/3-dibromo-17u-hydroxyprogesterone l7-acetate (the compound of Example 9) is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give 90;,1lfi-dibromo-lh-hydroxy-1,4-pregnadiene-3,20-dione 17-acetate.

EXAMPLE 21 1 7a-Hydroxy-1 ,4 ,9 (1 1 )-Pregnatriene-3,20-Dione I 7-Capr0ate 17a-hydroxy l,4,9(11) pregnatriene3,20'-dione (the compound of Example 13) is reacted with caproic acid in the presence of trifiuoroacetic anhydride and the resultant product isolated and purified in the manner described in Example 14 to give 17a-hydroxy-1,4,9(11)- pregnatriene-3,20-dione 17-caproate.

Alternatively, l7a-hydroxy-4,9( 11 -pregnadiene-3 ,20- dione 17-caproate (the compound of Example 2) is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15' to give 17a-hydroxy-1,4,9(11)-pregnatriene-3,20-dione 17-caproate.

EXAMPLE 22 1 7a-Hydroxy-1 ,4,9 (1 I -Pregnatriene-3,20-Dione 17-(fi-Cyclopentylpropionate) 9a,] 1 fi-Dich'loro-l 7a-Hydr0xy-1 ,4-Pregnadiene-3,20- Dione 1 7 p-Cyclopemyl propionate 17a-hydroxy l,4,9(11) pregnatriene 3,20-di0ne 17- 3cyclopentylpropionate), the compound of Example 22, is reacted with N-chlorosuccinimide in the presence of hydrogen chloride and lithium chloride in the manner described in Example 15 to give 9a,11 8-dichloro-17a-hydroxy-1,4-pregnadiene-3,20-dione 17 (/B-cyclopentylpropionate).

Alternatively, 9a,11fi dichloro 170t-hYdIOXy-4,9(11)- pregnadiene-3 ,20-dione 17 ,B-cyclopentylpropionate) the compound of Example 8, is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give :,11/3 dichloro 17a hydroxy-1,4-pregnadiene-3, ZO-dione 17-(fi-cyclopentylpropionate).

EXAMPLE 24 9a,] 1,8-Dichlor0-1 7u-H ydroxy-Z ,4 -Pregnadiene 3,20-Di0ne 17-Capr0ate 17a-hydroxy-1,4,9(11)-pre-gnatriene 3,20 dione 17- caproate (the compound of Example 21) is reacted with N-chlorosuccinimide in the presence of hydrogen chloride and lithium chloride in the manner of Example 15 to give 911,11,8-dichloro-l7ot-hydroXy-1,4-pregnadiene 3,20- dione 17-caproate.

Alternatively, 911,1 lfl-dichloro-l7a-hydroxy-4-pregnene- 3,20-dione 17-caproate (the compound of Example 7) is subjected to the action of a culture of Co rynebacterium simplex in the manner described in the alternative procedure of Example 15 to give '9a,11B-dichloro-17a-hydroxy-l,4-pregnadiene-3,ZO-dione 17-caproate.

EXAMPLE 25 9ot-I0d0-1 1 B-Bromo-I 7a-H ydroxy progesterone 1 7-A cctate To a solution of 1 g. of 17et-hydroxy-4,9'(1l)-pregnadiene-3,20-dione l7-acetate (prepared as described in Example 1), and 4 g. of lithium bromide in 50 ml. of acetic acid, there is added 620 mg. of N-iodosuccinimide followed by a solution of 220 mg. of hydrogen bromide in 4.5 m1. of acetic acid. The mixture is stirred at room temperature for one hour, then is poured into ice-water. A solid forms which is filtered, and then chromatographed on silica gel. The substance eluted with 30 ether in hexane is crystallized from acetone-hexane to give 9oc-10d0- 1lp-brorno-17a-hydroxyprogesterone 17-acetate.

EXAMPLE 26 17a-Hydroxy-19-N0r-4,9 (11 )-Pregnadiene-3,20-Dione (A) 115,17a-DIHYDROXY-19-NORPROGESTERONE A living culture of the organism Curvulariw lunata 1 5 (N.R.R.L. 2380) is rinsed from an agar slant under sterile conditions into a sterile medium having the following composition:

Percent Malt extract 5 Sucrose 1 Sodium nitrate 0.2 Potassium chloride 0.05 Magnesium sulfate heptahydrate 0.05 Ferrous sulfate heptahydrate 0.05 Dipotassium acid phosphate 0.1

Distilled water, to adjust to pH 7.0 with potassium hydroxide.

100 ml. of this medium is placed in each of several 300' ml. flasks. To each flask is added 50 mg. of l7a-hydroxy- 19-nor-progesterone dissolved in a smallvolume of acetone. The mixture is shaken for a period of 7 days at a room temperature of about 28 C. The contents of the flasks are then combined and extracted with several portions of ethylene dichloride using one-fifth the volume of the aqueous phase each time. The combined organic extracts are dried over sodium sulfate, filtered and concentrated in vacuo to a residue having a volume of 1-2 ml.

The ethylene dichloride residue is then placed on a chromatographic column consisting of silica gel, mixed with a small volume of methylene chloride. The column is developed with methylene chloride and the eluates are combined and concentrated in vacuo to a residue which is crystallized from acetone-hexane to give 116,17a-dihydroxy- 19-nor-progesterone.

(B) 17a-HYDROXY-19NOR 4,9(11)-PREGNADIENE- 3,20-DIONE Two grams of 11B,17a-dihydroxy-19-norprogesterone, prepared as described in Example 26A, in 50 ml. of pyridine is chilled to C., and there is added ml. of methanesulfonyl chloride dissolved in ml. of pyridine. The mixture is allowed to stand in the ice-box for 96 hours. A small amount of ice is added to the reaction mixture, and the solution is diluted with chloroform, washed with water, then with 5% hydrochloric acid, 5% sodium bicarbonate and finally again with water. The solution is dried over sodium sulfate, filtered, and concentrated in vacuo to a residue which is crystallized from methylene chloride-hexane to give 17a-hydroxy-19-nor-4,9(1l)- pregnadiene-3,20-dione.

EXAMPLE 27 90:,11/3-D ich loro-l 7a-Hydr0xy-1 9-Norprogester0ne 1.7-Acetate (A) 9o1,1lfi-DICHLORO-17a-HYDROXY-l9-NOR PRO GESTERONE One gram of 17a-hydroxy-19-nor-4,9(11)-pregnadiene-3,20-dione, prepared as in Example 26B, is dissolved in 30 ml. of carbon tetrachloride and at 20 C., there is added 220 mg. of chlorine dissolved in 30 ml. of carbon tetrachloride in the presence of 0.75 ml. of pyridine. The mixture is stirred at 20 C. for minutes, then allowed to warm to room temperature over a period of one-half hour. The solution is filtered and the filtrate concentrated in vacuo to a residue which is crystallized from acetone-hexane to give 90:,11 3 dichloro-l7a-hydroxy-19-norprogesterone.

(B) 9a,1lfi-DICHLORO-17a-HYDROXY-19-NOR- PROGESTERONE 17 ACETATE The 90;,115 dichloro-17a-hydroxy-19-norprogesterone of Example 27A is esterified with acetic acid in the presence of trifiuoroacetic anhydride and the resultant product isolated and purified in the manner described in Example 1 to give 90,1lfi-dichloro-l7a-hydroxy-l9-norprogesterone l7-acetate.

Similarly, by substituting other lower alkanoic acids such as propionic, butyric or caproic acid for acetic acid in the above procedure, the corresponding l7-esters are prepared, that is, the l7-propionate, 17-butyrate and 17- caproate of 9a,1lfl-dichloro-17u-hydroxy-l9-norprogesterone.

EXAMPLE 28 9a,]15-Dichloro-1 7oc-Hydr0xy-19-Norpr0gesterone 17-(fi-Cyclopentylpr pionale) 9:1,115 dichloro-lh-hydroxy-l9-norprogester0ne, the compound of Example 27A, is esterified by means of 13- cyclopentylpropionic acid and fl-cyclopentylpropionic anhydride under an atmosphere of nitrogen in the presence of p-toluenesulfonic acid in the manner of Example 3 to give 9a,1lp-dichloro-lh-hydroxy-19-norprogesterone 17- (,B-cyclopentylpropionate) EXAMPLE 29 9a-Br0m0-l 1,8-Chl0r0-1 7a-Hydr0xy-1 9-Norpr0gester0ne 1 7 A cetate (A) 9a-BROMO-llB-CHLORO-l7a-HYDROXY- IQ-NORPROGESTERONE 500 mg. of 17u-hydroxy-l9-nor-4,9(1l)-pregnadiene- 3,20-dione, the compound of Example 26B, is added to a solution of 220 mg. of N-bromoacetamide and 2 g. of lithium chloride in 20 ml. of glacial acetic acid. The mixture is stirred at room temperature for about 30 minutes then poured into ice-water. A solid product separates which is filtered, washed well with water and crystallized from methylene chloride-pentane to give 9abromo-l 1,8 chloro-17a-hydroxy-19-norprogesterone.

(B) 9a-BROMO-l'lfi-CHLORO-l'l'a-HYDROXY-LLQ- NORPROGESTERONE l7-ACETATE In the manner described in Example 1, 9a-bromo-l1fichloro-l7a-hydroxy-19-norprogesterone, the compound of Example 29A, is esterified with acetic acid in the presence of trifluoroacetic anhydride to give 9a-bromo-llflchloro-17a-hydroxy19-norprogesterone l7-acetate.

In a similar fashion, other lower alkanoic acid esters of the compound of Example 29A are prepared by substituting suitable acids such as propionic, butyric and caproic for acetic acid in the above procedure.

EXAMPLE 3O 9a-Br0mo-1 1 B-Fluoro-I 7a-Hydr0xy-l 9-N0rpr0gesterone 1 7 A cetate (A) 9a-BROMO-1lfl-FLUORO17a-HYDROXY-l9- NORPROGESTERONE In the manner described in Example 6, 250 mg. of 17a-hydroxy-19-nor-4,9(11)-pregnadiene-3,20-dione (the compound of Example 26B) is reacted with mg. of N-bromoacetamide in 50 ml. of diethylacetic acid and a solution of 250 mg. of hydrogen fluoride in 2 ml. of a chloroform-tetrahydrofuran mixture, and the resultant product isolated and purified to give 9a-bI'0InO-llB-fllloro-l7a-hydroxy-l9-norprogesterone.

(B) 9oz-BROMO-1lfi-FIJUORO-17a-HYDROXY-19- NORPROGESTERONE 17-ACETATE 17 EXAMPLE 31 9a-Iodo-l 1,8-Chlro-1 7a-Hydroxy-19-N0rprogesterone 1 7-A cetate (A) 9uz-IODO-11B-CHLORO-l'Ta-HYDROXY-19- N ORPROG-EJSTERONE In the manner described in Example 10, 17a-hydroxy- 19-nor-4,9(11)-pregnadiene-3,20-dione (the compound of Example 26B), is reacted with iodine monochloride in tetrahydrofuran and the resultant product isolated and purified to give 9u-iodo-1lit-chloro-17a-hydroxy-19-nor progesterone.

(B) 9wIODO-11.13-CHLORO-I'icrHYDROXY-IQ- NORPROGESELERONE 17-ACETATE The 17u-hydroxy-19-n0rprogesterone of Example 31A is esterified to the corresponding 17-acetoxy compound by means of acetic acid in the presence of trifiuoroacetic anhydride in the manner described in Example 1 to give 906 iodo-l 1 8-chloro-17ot-hydroxy-19-norprogesterone 17 acetate.

EXAMPLE 32 1 7rx-Methyl-4,9 (.1 1 )-Pregnadiene-3,20-Dione (A) 17MMETHYL-2l-HYDROXY=4=,9(11) PREGNADIENE 320-DIONE 21-METHANESULF'ONATE A solution of g. of 17a-methyl-11,8,21-dihydroxy-4- pregnene-3,20-dione in 100 ml. of pyridine is chilled to 0 C. and a solution of ml. of methanesulfonyl chloride in 20 ml. of chloroform is added dropwise. The mixture is allowed to stand in the ice box for 100 hours, then a little ice is added and the solution diluted with chloroform. The organic solution is washed with Water, 5% hydrochloric acid, 5% aqueous sodium bicarbonate, and again with water. The solution is dried over magnesium sulfate, filtered and concentrated in vacuo to a residue which is crystallized twice from acetone-hexane to give 17 ot-methyl-21-hydroxy-4,9( 11)-pregnadiene-3,20- dione 21-methanesulfonate.

(B) 17a-METHYL4,9(11)-PREGNADIENE-3,20-DIONE To a solution of 2.0 g. of 17a-methyl-21-hydroxy-4, 9(1'1)-pregnadiene-3,20-dione ZI-methanesulfonate (the compound of Example 32A), in 30 ml. of acetic acid is added 4.0 g. of sodium iodide. The mixture is heated on the steam-bath for 30 minutes, then cooled and decolorized by the addition of aqueous sodium bisulfite solution. The solution is then poured into ice water. A precipitate forms which is filtered, Washed with Water, and crystallized from acetone-hexane to give 17a methy1- 4,9 1 1)-pregnadiene-3,20-dione.

EXAMPLE 3 3 9a,] 1 fi-Dich l0r0-1 7a-Methylpr0gesterone One gram of 17/x-methyl-4,9(l1)-pregnadiene-3,20-dione, the compound of Example 32, is reacted with N- chlorosuccinimide (430 mg.) in the presence of hydrogen chloride and lithium chloride in the manner of Example 4. The resultant product is isolated and purified in the described manner to give 9a,1lfi-dichloro-17a-methylprogesterone.

EXAMPLE 34 9ot-Br0m0-1 1 fl-Fluoro-l 7 a-M ethy [progesterone In the manner described in Example 6, one gram of 17ot-methy1-4,9(11)-pregnadiene-3,20-dione, the compound of Example 32, is reacted with N-bromoacetamide (450 mg.) and hydrogen fluoride in diethylacetic acid. The isolated product is crystallized from methylene chloride-pentane to give 9a-bromo-11fi-fluoro-17rx-methylprogesterone.

EXAMPLE 35 9oc-BI0I7'tO-1 1 fi-Chloro-l 7 a-M ethy l progesterone In the manner described in Example 5, 17a-methyl-4,

EXAMPLE 36 17a-Methyl-1,4,9(11 -Pregnatriene-3,20-Dione In the manner described in the alternative procedure of Example 15, 17a-methyl-4,9(11)-pregnadiene-3,20-di one is subjected to the action of a culture of Corynebacterium simplex and the resultant product isolated and puri fied to give 17a-methyl-1,4,9(1l)-pregnatriene-3,20-dione.

EXAMPLE 3 7 :,11fl-DiChIOfO-1 7 a-M ethyl-1 ,4 -Pregnadiene-3,20-D tone The l7ot-methylpregnatriene of Example 36 is reacted With N-chlorosuccinirnide in the presence of hydrogen chloride and lithium chloride in the manner described in Example 4, and the resultant product isolated and purified in the described manner to give 90,11fi-dlClJl0lO-17otmethyl-1,4-pregnadiene-3,ZO-dione.

EXAMPLE 38 The 17a-methylpregnatriene of Example 36 is reacted With N-bromoacetarnide and hydrogen fluoride in diethylacetic acid and the resultant product isolated and purified in the manner of Example 6 to give 9oc-b101'I10-11fl-fltt010- 17u-1'I16il'lYl-1 ,4-pregnadiene-3 ,ZO-dione.

EXAMPLE 39 9a-Chl0r0-J 1 [i-Flzzoro-I 7ot-Methyl-1,4-Pregnadiene- 3 ,2 O-Dione The l7a-methylpregnatriene of Example 36 is reacted with N-chlorosuccinimide and hydrogen fluoride and the resultant product isolated and purified in the described manner of Example 11 to give 9ot-chloro-lLB-fluoro-17amethyl-1,4-pregnadiene-3,20-dione.

EXAMPLE 4O 90:,11/3-Dibl0h10-1 7 (PM ethyl-1 ,4-Pregnadi ene3,20-Di0ne In the manner described in Example 9, 17a-methyl-1,4, 9(11)-pregnatriene-3,ZO-dione, the compound of Example 36, is brominated by means of N-bromoacetamide in the presence of hydrogen bromide and lithium bromide and the product isolated and purified to give 90,l15-dibromo-l7a-methyl-1,4-pregnadiene-3,ZO-dione.

EXAMPLE 41 9oc-I0tl'0-1 lfl-Chloro-I 7 a-M ethylprogesterone In the manner described in Example 10, 500 mg. of 17a-methyl-4,9(11)-pregnadiene 3,20 dione, the compound of Example 32, is reacted With 250 mg. of iodine monochloride and the resultant product isolated and purified to give 9a-iodo-1Iii-chloro-17ot-methylprogesterone.

EXAMPLE 42 J 7a-Br0m0-4,9 (J1 -Pregnaa' iene-3 ,2 O-Dione (A) 17a.-BROMO-11B-I-IYDROXYPROGES-TERONE 17u-bromoprogesterone is subjected to the action of a culture of Curvularia lzmata in the manner described in Example 26A and the resultant product isolated in the described manner and crystallized twice from ethylacetate to give 17ot-bromo-1lfl-hydroxyprogesterone.

(B) z-BROMO-4,9 (11) -PREGNADIENE-3,20-DIONE A mixture of 2 g. of 17a-bromo-1lfi-hydroxyprogesterone, prepared as in Example 42A, and 4 g. of dry lithium bromide in 50 ml. of glacial acetic acid is refluxed under anhydrous conditions for 30 minutes. The solution is then cooled and poured into ice water. A Solid precipitates which is filtered washed well with water and recrystallized twice from acetone-hexane to give l7a-bromo-4, 9(11)-pregnadiene-3,20-dione.

EXAMPLE 43 9a,] 1/3-Dichl0r0-1 7u-Bromoprogesterone One gram of 17a-bromo-4,9(l l)-pregnadiene-3,20-dione, the compound of Example 42, is reacted With 370 mg. of N-chlorosuccinimide in the presence of hydrogen chloride and lithium chloride in the manner described in Example 4. The resultant product is isolated in the described manner and crystallized from methylene chloridehexane to give 911,1lB-dichloro-l7a-bromoprogesterone.

EXAMPLE 44 I 9a,] 7a-Dibr0m0-Z lp-Chloroprogesterone 500 mg. of the 17a-bromopregnadiene of Example 42 is reacted with 190 mg. of N-bromoacetamide in the presence of hydrogen chloride and lithium chloride in the manner described in Example 5. The resultant product is isolated in the described manner and crystallized from acetone-hexane to give 9a,17a-dibromo-llfl-chloroprogesterone.

EXAMPLE 45 500 mg. of the 17o -bromopregnadiene of Example 42 is reacted with 180 mg. of N-chlorosuccinimide in the presence of hydrogen fluoride in the manner described in Example 11. The resultant crude product is chromatographed on silica gel and the material eluted with 30% ether in hexane is crystallized twice from methylene chloride-pentane to give 9a-chloro-llfl-fiuoro-lh-bromoprogesterone.

EXAMPLE 46 1 7OL-BI'OI?ZO-1,4,9 I 1 )-Prcgnatriene-3,20-Dione The 17or-brorno-4,9(11)-pregnadiene-3,20-dione of Example 42 is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 and the resultant product isolated and purified in the described manner to give 17abromo-1,4,9 l l pregnatriene-3,20-dione.

EXAMPLE 47 9a,11fi-Dicl1l0r0-1 7 ot-Brom -1 ,4 Pregnadien-3 ,2 O-Dione The 17a-bromopregnatriene of Example 46 is reacted with N-chlorosuccinimide in the presence of hydrogen chloride and lithium chloride in the manner described in Example 4. The resultant product is isolated and purified in the described manner to give 90:,11B-diChIOIO-17otbromo-l,4-pregnadiene-3,20-dione.

Alternatively, the compound of this example is prepared from 9a,1LB-dichloro-17a-bromoprogesterone, the compound of Example 43, with the aid of a culture of Corynebacterium simplex in the manner of the alternative procedure of Example 15.

EXAMPLE 48 9a,]7oc-Dibl'0l710-1 1B-Chloro-I,4-Pregnadiene-3,20-Dione 17a-bIOI110-1,4,9( 1 1 -pregnatriene-3,20-dione, prepared as in Example 42, is reacted with lithium chloride, N- bromoacetamide and hydrogen chloride, and the resultant product isolated and purified in the manner described in Example 5 to give 9a,17a-dibromo-1lfl-chlorol,4-pregnadiene-3,20-dione.

Alternatively, the compound of this example is prepared from 9oz,l7oc-dibromo-llB-chloroprogesterone, the compound of Example 44, With the aid of a culture of Corylzebacterimn simplex in the manner described in the alternative procedure of Example 15.

20 EXAMPLE 49 9a,]7a-Dibr0m0-11fl-Fluoro-I,4-Pregnadiene-3,20-Dione In the manner described in Example 6, 17a-bromo- 1,4,9(l1)pregnatriene-3,20-dione, the compound of Example 42, is reacted with N-bromoacetamide and hydrogen fluoride in diethylacetic acid. The resultant product is isolated and purified in the described manner to give 9a,17or-dibromo-1 lfl-fluoro-1,4-pregnadiene-3,20-dione.

EXAMPLE 5 0 9a,]1B,17a-Tribr0m0-1,4-Pregnadiene-3,20-Di0ne In the manner described in Example 9, l7a-bromo- 1,4,9(l1)-pregnatriene-3,20-dione (the compound of Example 42) is reacted with lithium bromide, N-bromoacetamide and hydrogen bromide in acetic acid and the resultant product isolated and purified to give 9 x,1l 3,1704- tribromo-l,4-pregnadiene-3,20-dione.

EXAMPLE 5 1 1 7 oc-Ch loroprogesterone (A) 17a-HYDROXY-5a,GBJ'Ya-TRICHLOROaXLLO- PREGNANE-ZO-ONE S-ACETATE The requisite starting compound, 5,17(20)-pregnadiene-3fl,20-diol diacetate, is prepared as follows.

To a solution of 5 g. of 3B-hydroxy-5-pregnene-20-one 3-acetate in 250 ml. of acetic anhydride there is added 1.5 g. of p-toluenesulfonic acid. The mixture is slowly concentrated to about one-fourth the original volume by distillation over a six hour period. The remaining solvent is then distilled in vacuo. The resulting residue is taken up in ether, washed with water, then with aqueous sodium bicarbonate solution and then again with water. The ether solution is dried over magnesium sulfate, filtered, then passed through a column of Florisil in ether. The eluate is concentrated to a residue which crystallizes upon the addition of hexane. The crystalline material is filtered and air dried to give 5,17(20)-pregnadione- 35,20-diol diacetate.

The 5-pregnene diacetate prepared as described above is dissolved in ml. of carbon tetrachloride, 2 ml. of pyridine is added, the solution chilled to -20" C. and then there is added dropwise a solution of 1 g. of chlorine dissolved in 8.4 ml. of carbon tetrachloride. The mixture is stirred at 20 C. for one-half hour, then allowed to come to room temperature. The solution is diluted with methylene chloride, washed with water, 5% sodium bicarbonate, 5% hydrochloric acid and again with water, dried over magnesium sulfate, then concentrated in Vacuo to a residue which is crystallized from acetone-hexane to give 3B-hydroxy-5a,6;8,l7atrichloroallopregane-ZO-one 3-acetate.

(B) 3B-I-IYDROXY-1 7a-CHLORO-5-PREGNENE-ZO-ONE 8-ACETATE A solution of 2 g. of the compound of Example 51A in 50 ml. of methyl ethyl ketone containing 2 g. of sodium iodide is refluxed under nitrogen for 2 hours, then poured into cold water. A precipitate forms which is filtered, Washed with water and crystallized from ethyl acetate to give 3/3-hydroxy-17a-chloro-S-pregnene-ZO-one 3-acetate.

(C) 17a-CHLOROPROGESTERONE 3B-hydroxy 17oz chlOro-S-pregnene-ZO-One 3-acetate, prepared as in Example 51B, is subjected to the action of a culture of Flavobacterium dehydrogenans (Rutgers Collection No. as follows. The culture of the organism is prepared by propagating it in a nutrient agar medium at 30 C. for 24 to 72 hours. During incubation, the inoculated tube is exposed to light with the resultant development of a yellow pigment characteristic of the species. The developed culture is rinsed from an agar slant under sterile conditions into a sterile medium of pH 6.8 and having the following composition:

This culture medium has previously been autoclaved, at 15 lb. pressure, for twenty minutes to obtain aseptic conditions, and cooled. The variant is grown in the medium under constant illumination, using the visible range of the spectrum. The incubation temperature is maintained at about 33 C. and is conducted under aerobic conditions. Aeration is accomplished by agitation and/or blowing air through the culture medium.

After the organism has grown for 12 to 24 hours (or longer, if desired), 100 ml. of the growing culture are introduced into each of ten flasks, and to each flask are added 200 mg. of 3,8-hydroxy l7u-chloro-5-pregnene-20- one 3-acetate dissolved in a minimum volume of ethanol. The reaction mixtures are then shaken at 30 C. for 12 to 72 hours. The reaction is stopped when paper chromatography indicates that there is no more starting material.

The contents of the flasks are combined and extracted with methylene chloride. The extracts are concentrated and the residue is crystallized from acetone-hexane yield ing 17u-chloroprogesterone.

EXAMPLE 5 2 1 7a-Chl0ro-4,9 (11 )-Pregnadiene-3,20-Dine (A) 1lfl-HYDROXY-l7a-CI-ILOROPROGESTERONE 17u-chloroprogesterone, prepared as described in Example 51, is subjected to the action of a culture of Curvularia lunam and the resultant product isolated in the manner described in Example 26A to give a residue which is crystallized from ethyl acetate to give 1113- hydroxy-17a-chloroprogesterone.

( B) 17a-CHLORO-4,9 (11)-PREGNADIENE3,20-DIONE 1lfi-hydroxy-l7a-chloroprogesterone, prepared as in Example 52A, is reacted with lithium bromide in glacial acetic acid in the manner described in Example 42B and the resultant product isolated and purified to give 17a-chloro-4,9(l1)-pregnadiene-3,20-dione.

EXAMPLE 5 3 9a,]1fl-1 7u-Trichloroprogesterone One gram of l7a-chloro-4,9(1l)-pregnadiene-3,20- dione, prepared as in Example 52B, is reacted with 390' mg. of N-chlorosuccinimide in the presence of hydrogen chloride and lithium chloride in the manner described in Example 4. The resultant product is isolated in the described manner and crystallized from acetone-hexane to give 9a,1lfi,17a-trichloroprogesterone.

EXAMPLE 54 9ot-Br0m0-1 1,8,1 7u-Dichl0roprogesterone One gram of l7a-chloro 4,9(1l)-pregnadiene-3,20 dione, the compound of Example 523, is reacted with 400 mg. of N-bromoacetamide in the presence of hydrogen chloride and lithium chloride according to the procedure of Example 5. The resultant product is isolated in the described manner and crystallized twice from methylene chloride-pentane to give 9a-bromo-11/3,17adichloroprogesterone.

EXAMPLE 55 9a-Bnomo-1 1 ,B-Fluoro-I 7 a-Chloroprogesterone In the manner described in Example 6, the 4,9(1l)- pregnadiene of Example 52B is reacted with N-bromoacetamide and hydrogen fluoride in diethyl acetic acid and the resultant product isolated and purified. to give 9abromo-l Iii-fluoro-17a-chlor0progesterone.

22 EXAMPLE 56 In the manner described in Example 11, the 4,9(11)- pregnadiene of Example 52B is reacted with N-chlorosuccinimide and hydrogen fluoride and the resultant product isolated and purified to give ,17ot-(li0h101O-11fifluoroprogesterone.

EXAMPLE 57 1 7 oc-Cll Zora-1 ,4,9 (1 1 )-Pregnatriene-3,20-Dione 17a-Chl01O-4,9( ll)-pregnadiene-3,20-dione, the compound of Example 52B, is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15. The resultant product is isolated and purified in the described manner to give 17a-chloro-1,4,9(ll)-pregnatriene-3,20- dione.

EXAMPLE 58 901,1 15,1 7a-Trichl0ro-1 ,4-Pregnad iene-3,20-Di0ne In the manner described in Example 4, 17a-Chl010- 1,4,9(ll)-pregnatriene-3,ZO-dione, the compound of Example 57, is reacted with N-chlorosuccinimide in the presence of hydrogen chloride and lithium chloride and the resultant product isolated and purified to give 9a,l1fi, 17a-trichloro-l,4-pregnadiene-3,20-dione.

Alternatively, the compound of this example is prepared from 9a,l1B,17a-trichloroprogesterone, the compound of Example 53, with the aid of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15.

EXAMPLE 59 Qa-Bromo-l 1B,].7a-Dichlore-1,4-Pregnadiene-3,20-Dione In the manner described in Example 5, the 17erchloropregnatriene of Example 57 is reacted with N-bromoacetamide in the presence of hydrogen chloride and lithium chloride, and the resultant product isolated and purified to give 9a-brorno-l1B,17a-dichloro-l,4 pregnadiene -3,20-dione.

Alternatively, 9a bromo llfl,l7a-dichloroprogesterone, prepared as in Example 54, is subjected to the action of a culture of Corynebacterium simplex in the manner described in the alternative procedure of Example 15 to give 9ot-bromo-11B,17a-dichloro-1,4-pregnadiene-3,20- dione.

EXAMPLE 60 9a-Brom0-1 1 fi-Fluora-l 7a-Chl0r0-1,4-Pregnadiene- 3,20-Di0ne In the manner described in the alternative procedure of Example 15, 9a-bromo-llit-fluoro-17a-chloroprogesterone, the compound of Example 55, is subjected to the action of a culture of Corynebacterium simplex and the resultant product isolated and purified to give 9a-bromo- 11 3-fluorol7u-chloro-1,4-pregnadiene-3,20-dione.

EXAMPLE K61 9a,]7a-Dichl0ra-11j3-Flrz0r0-l,4-Pregnadiene-3,20-Diane In the manner described in the alternative procedure of Example 15, 9a,17a-dichloro-1lp-fluoroprogesterone, is subjected to the action of a culture of Corynebacterium simplex and the resultant product isolated and purified to give 9a,17ot-dichloro-1lit-fluoro-1,4-pregnadiene-3,20- dione.

EXAMPLE 62 In the manner described in Example 9, 17x-Ch101'0- 1,4,9(11)-pregnatriene-3,20-dione, is reacted with N-bromoacetamide in the presence of lithium bromide and hy- 23 drogen bromide and the resultant product isolated and purified to give 9a,llfi-dibromo-l7a-chloro-l,4-pregnadiene-3,20-dione.

EXAMPLE 63 In the manner described in the alternative procedure of Example 15 9oc-iodo-llfl-chloro-l7a-methylprogesterone (the compound of Example 41) is subjected to the action of a culture of Corynebacterium simplex and the resultant product isolated and purified to give 9oc-i0d0- 1 lfi-ch'loro-l 7a-methyl-1,4 pregnadiene-3 ,20-dione.

EXAMPLE 64 9a,] 1 fi-Dibrom -1 7 oc-M ethyl progesterone 17 a-methyl-4,9( 1 1)-pregnadiene-3,20-di0ne (the compound of Example 32B) is reacted with N-bromoacetamide, lithium bromide. and hydrogen bromide and the resultant product isolated and purified in the manner described in Example 9 to give 904,11B-dibromo-17a-methylprogesterone.

EXAMPLE 65 17oc-bIOII10-4,9 l 1 )-pregnadiene-3,20-dione (the compound of Example 42B) is reacted with lithium bromide, hydrogen bromide and N-iodo-succinimide in the manner described in Example 25. The resultant product is isolated and purified in the described manner to give 9oc-iOdO-1 lfl,17a-dibromoprogesterone.

In the manner of Example 9, 170t-b1'OIl'10-4,9(11)- pregnadiene-3,20-dione (the compound of Example 42B) is reacted with N-bromoacetamide, lithium bromide and hydrogen bromide, and the resultant product isolated and purified to give 911,11B,17a-tribromoprogestr0ne.

EXAMPLE 67 9oc-l ado-1 1 6,1 7 u-D ibromo-J ,4-Pregnadiene-3,20-Di0ne 9a-Iodo-l1 8,17a-dibromoprogesterone (the compound of Example 65) is subjected to the action of a culture of Corynebacterium simplex and the resultant product isolated and purified in the manner described in the alternative procedure of Example to give 9u-iodo-11fi,17ocdibromo-1,4-pregnadiene-3,20-di0ne.

EXAMPLE 68 9a,]1fl-Dibr0mo-17a-Hydr0xy-19-N0rpr0gesterone 17-Acetate (A) 9a,11fi-DIBROMO-17a-HYDROXY-19- NORPROGESTERONE To a solution of 1 g. of l7u-hydroxy-4,9(11)-19- norpregnadiene-3,20-dione (the compound of Example 26B) and 4 g. of lithium bromide in 50 ml. of acetic acid is added 420 mg. of N-brornoacetamide. The mixture is stirred at room temperature for one hour, then is poured into ice-water. A solid separates which is filtered, Washed with water, and crystallized from acetone-hexane to give 9a,l1,8-dibromo-17a-hydroxy-l9-norprogester0ne.

(B) 9a,1lfi-DIBROMO-17a-HYDROXY-19-NOR- PROGESTERONE 17-ACETATE The dibrom0-l7a-progesterone of Example 68A is esterified in the manner described in Example 1 to give 9a,1lfl-dibromo-lh-hydroxy 19 norprogesterone 17- acetate.

EXAMPLE 69 9a,] 1 9-Dichl0r0-1 7a-H ydroxy progesterone 1 7-A cetate (A) 9a,115=DICHLOR0-17a-HYDROXYPROGESTERONE One gram of 17a-l1ydroxy-4,9(l1)-pregnadiene-3,2O-

24 dione is reacted with 220 mg. of chlorine in carbon tetrachloride in the presence of pyridine in the manner described in the alternative procedure of Example 4 to give 9a,llfi-dichloro-l7a-hydroxyprogesterone.

xfjgi? 238 m (B) 9e,11,B-DI'CHLORO-17a-HYDROXYPROGESTERONE 17-ACETATE In the manner described in Example 14, :,1lB-dichloro-17a-hydroxyprogesterone (the compound of Example 69A) is esterified by means of trifluoroacetic anhydride in acetic acid to give 9a,11fi-dichloro-17a-hydroxyprogesterone 17-acetate.

In similar manner, by substituting other lower alkanoic acids, such as butyric and valeric, for acetic acid in the esterification procedure, other lower alkanoyl esters are prepared such as the 17-butyrate and 13-valerate of 911,1 1 fl-dichloro-l7a-hydroxyprogesterone.

Alternatively, the compound of this example is prepared according to the following procedure.

(C) 9a,11B-DIC'HLORO-17a-HYDROXYPROGESTERONE 17-ACETATE One gram of 17a-hydroxy-4,9(11)-pregnadiene-3,20- dione 17-acetate (the compound of Example 1) and 850 mg. of p-iodotoluene dichloride are dissolved in 20 ml. of methylene chloride. The solution is stirred at room temperature for 5 hours, then concentrated in vacuo to a residue which is crystallized from acetone-hexane to give 9a,1lfl-dichloro-17a-hydr0xyprogesterone 17-acetate.

EXAMPLE 7O 9oc-Br0m0-1 1 fi-Ch lore-1 7oc-H ydroxy progesterone 1 7 -A ceate (A) 9a-BROMO-11fl-CHLORO-17a-HYDROXY- PROGESTERONE One gram of 17u-hydroxy-4,9(ll)-pregnadiene-3,20- dione and 4 g. of lithium chloride are dissolved in 50 ml. of glacial acetic acid. There is first added 420 mg. of N-bromoacetamide and then 111 mg. of hydrogen chloride in 40 ml. of acetic acid. The mixture is stirred at room temperature for about 30 minutes then is poured into ice-water with stirring and the resultant solid is filtered, washed with water, and crystallized from acetonehexane to give 9a-bromo-1lfi-chloro-17a-hydroxyprogesterone,

A3552 241 mm (B) 9a-BROM0-11B-CHLORO-17a-HYDROXYPROGE- STERONE 17-ACETATE The 17a-hydroxy compound of Example 70A is esteritied with acetic acid in the presence of trifluoroacetic anhydride in the manner of Example 14 to give 9a-bromo- 1lfi-chloro-17u-hydroxyprogesterone 17-acetate.

In similar fashion, by substituting other lower alkanoic acids such as butyric and valeric for acetic acid in the esterification procedure, other lower alkanoyl esters are prepared, such as the 17-butyrate and 17-valerate of 9abromo-l lfl-chlorol7a-hydroxyprogesterone.

EXAMPLE 71 17a-Hydr0xy-1,9:(11 )Allopregnadicne-3,20-Dione 17-Acetale (A) 1113,17a-DIHYDROXY-1-ALLOPREGNENE-3,20- DIONE 17'ACETATE In the manner described in Example 26A, 17a-hydroxy- 1-allopregnene-3,20-dione 17-acetate is subjected to the action of a culture of the organism Curvularia lunata. The resultant product is isolated and purified in the described manner to give 1118,17a-dihydroxy-l-allopregnene- 3,20-dione 17-acetate.

(B) 17aHYDROXY-1,9(11)-ALLOPREGNADIENE-3,20- DIONE 17-ACETATE The llfl-hydroxy allopregnene of Example 71A is 25 reacted with methanesulfonyl chloride in pyridine and the resultant product isolated and purified in the manner described in Example 2613 to give 17u-hydroxy-1,9(l1)-a1lopregnadiene-3,20-dione 17-acetate.

EXAMPLE 72 9a,11fi-Dichloro-I7a-Hydroxy-1-Allopregnen e-3,20- Dione 1 7-A cetate 17a hydroxy l,9(11)-allopregnadiene-3,20-dione 17- acetate (the compound of Example 71) is chlorinated with chlorine in carbon tetrachloride in the presence of pyridine and the resultant product isolated and purified in the manner described in Example 27A to give 90:,11fldichloro-17ahydroxy-1-allopregnene-3,20-dione 17-acetate.

EXAMPLE 73 9u-Br0mo-1 1 B-ChZOro-l 7 a-H ydroxy-I -A llopregnen- 3,20-Dione 17-Acetate 17a hydroxy 1,9(11)-allopregnadiene-3,20-dione l7- acetate (the compound of Example 71) is reacted with 4 g. of lithium chloride, 420 mg. of N-bromoacetamide and anhydrous hydrogen chloride in acetic acid in the manner of Example 5. The resultant product is isolated and purified in the described manner to give 9u-bromo-11B- Chloro 17a hydroxy-1-al-lopregnene-3,20-dione 17-acetate.

EXAMPLE 74 9a,] 1 B-Dichloro-I 7a-Hydr0xy-1,4-Pregnadiene- 3,20-Dione 17-Acetwte 1 7a-Methyl-1 -A llopregnene-3,20-Dine (A) 17a-MEIHYLALLOPREGNANE-3,20'DION E A solution of 1.0 g. of 17a-methylprogesterone in ml. of tetrahydrofuran is added in a slow stream with stirring to a solution of 400 mg. of lithium metal in 200 ml. of liquid ammonia chilled to about 70 C. in a Dry Ice-acetone 'bath. The reaction mixture is allowed to stir for 2 minutes, then sufficient solid ammonium chloride is added to discharge the blue color. The solution is allowed to evaporate to a residue. Water is added to the residue and the mixture extracted with ether. The extracts are combined and washed with water, dried over magnesium sulfate, filtered, and evaporated to a residue which is crystallized from acetone-hexane to give 170:- methylallopregnane-3,ZO dione.

(B) 2'-BROMO-17aJHETHYLALLOPREGNANE-3,20-DIONE To a solution of 2.0 g. of the allopregnane of Example 75A in 100 ml. of acetic acid is added a drop of hydrogen bromide in acetic acid solution, followed by the dropwise addition, with stirring, of a solution of 1.0 g. of ibromine in 36 ml. of acetic acid. The solution is stirred for a few minutes until the bromine color is discharged, and then diluted with cold water. A solid precipitate forms which is filtered and crystallized from acetone-ether to give 2-bromo-17a-methylal1opregnane- 3,20-dione.

(C) 17a-METHYL-1-ALLOPREGNENE-3,20-DIONE 1.5 grams of the Z-bromo compound of Example 75B is refluxed in m1. of collidine for one hour. The mixture is diluted with chloroform and the layers are separated. The organic layer is washed with 5% aqueous hydrochloric acid and water, dried over magnesium sulfate, filtered, and evaporated to a residue which is crystallized from acetone-hexane to give l7a-methyl-1-allopregnene-3,20-dione.

EXAMPLE 76 1 7 u-M ethyl-1 ,9 (11 -A llopregnadiene-3,20-Di0ne (A) llfl-HYDRQXY-l7a-METHYL-1-ALLOPREGNENE 3,20-DIONE 17a-Methyl-1-allopregnene-3,20-dione (the compound of Example 75) (2.0 g.) is llfi-hydroxylated by fermentation with Curvularia lunata (N.-R. R.L. 2380) in the manner of Example 26. The product is isolated in the described manner and crystallized (from acetone-hexane to give 1lfl-hydroxy-lh-methyl-1-allopregnene-3,20- dione.

(B) 17a-METHYL-l,9 (11) -ALLOPREGNADIENE- 3,20-DIONE The lip-hydroxy compound of Example 76A (2.0 g.) is dehydrated by treatment with methanesul-fonyl chloride in pyridine in the manner described in Example 2613. The product is isolated in the described manner and crystallized from acetone-hexane to give 17u-methyl-1,9(11)- allopregnadiene-3,20-dione.

EXAMPLE 77 :,11B-Di0hl0l0-1 7 a-M ethyl-1 -A l lopregnene-3,20-Dione 17 a-Methyl-1,9 1 l allopregnadiene-3,ZO-dione (the compound of Example 76) is chlorinated with chlorine in carbon tetrachloride in the presence of pyridine in the manner of Example 27A. The resultant product is isolated and purified in the described manner to give 911,1 118- dichloro-17a-Methyl-l-allopregn'ene-3,20-dione.

EXAMPLE 78 9a-Br0mo-11 8-Chlor0-1 7a-Methyl-1-All0pregnene-3,20- Dione l7a-methyl-l,9(1 l )-allopregnadiene-3 ,20-dione (the compound of Example 76) is reacted with 4 g. of lithium chloride, 420 mg. of N-bromo-aceta-mide and anhydrous hydrogen chloride in acetic acid in the manner of Example 5. The resultant product is isolated and purified in the described manner to give 9a-bromo-l lB-ChlOIO-17ocmethyl-l-allopregnene-3,20-dione.

EXAMPLE 79 90:,11B-Di0hl070-1 7 a-M ethyl-1 ,4-Pregnadiene-3,20- Dione One gram of 9a,llfl-dichloro-17m-methyl-1-allopregnene-3,20-dione (the compound of Example 78) is reacted with 0.3 g. of selenium dioxide in 50 ml. of t-butyl alcohol and 1 ml. of acetic acid under nitrogen in the manner described in Example 74. The resultant product is isolated and purified in the described manner to give 9oz,l IB-dichloro-lh-methyl- 1,4-pregnadiene-3,20-dione.

EXAMPLE 80 9a-l1,B-Dichlorw17a-Hydr0xy-All0pregnene-3,20-Di0ne (A) 17a-HYDROXY-1,9 (11)-ALLOPREGNADIENE- aaomoun 1 A solution of 1 g. of 17a-hydroxy-1,9(11)-allopregnadiene-3,20-dione l7-acetate (the compound of Example 71) in ml. of a 3:1 methanol-chloroform mixture is chilled in ice and there is added dropwise 52 ml. of 0.1 Normal sodium hydroxide solution (1 equivalent). The reaction mixture is stirred at 0 C. for 10 minutes, then diluted with Water and extracted with methylene chloride. The organic layer is washed with water, dried over magnesium sulfate, filtered and evaporated to a residue which is crystallized from acetone-hexane to give Net-hydroxy- 1,9 l l -allopregnadiene-3,20-dione.

(B) 901,1lflDICHLORO-l7a-HYDROXYl-ALLO- PREGNENE3,20-DIONE 17a-hydr0xy-1,9(11)-allopregnadiene-3,ZO-dioue (prepared as in Example 80A) is chlorinated with chlorine in carbon tetrachloride in the presence of pyridine and the resultant product isolated and purified in the manner described in Example 27 to give 9a,11fl-dichloro-17a-hydroxy-1-allopregnene-3,20-dione.

Alternatively, the compound of this example is prepared from 9a, 1lfi-dichloro-lh-hydroxy-1-allopregnene-3,20- dione (the compound of Example 72) by reaction with one equivalent of 0.1 Normal sodium hydroxide solution in the manner described in Example 80A to give 90:,11fldichloro-l7ot-hydroxy-1-allopregnen'e-3,20-dione.

EXAMPLE 81 9a,IJ/3-Dichl0r0-1 7u-Hydr0xy-1-All0pregnene-3,20- Dione 17-Capr0ate In the manner described in Example 2, 90,l1B-Dichloro-17a-hydroxy-l-allopregnene-3,20-dione (the compound of Example 80) is reacted with caproic acid and trifluoroacetic anhydride to give 9a,11fi-dichloro-17a-hydroxy-l-allopregnene-3,20-dione 17-caproate.

In similar fashion, other lower alkanoic acid esters of the compound of Example are prepared by substituting suitable acids such as propionic and butyric for caproic acid in the above procedure to give the 17-propionate and 17-butyrate respectively of 9oc,1lfi-dichloro-lh-hydroxy- 1-allopregnene-3,20-dione.

We claim:

1. A compound selected from the group consisting of 17oz-R9( l1)-dehydnoprogesterone, 17oc-R-1,9 1 1 )-bisdehydroprogesterone and 17a-R-1lfi-hydroxyprogesterone wherein -R is a halogen having an atomic weight greater than 19 and less than 126.

2. 17a-Bromo-9 11)-dehydroprogesterone.

3. 17a-Chloro-9 1 1 dehydroprogesterone.

4. 1 1,B-Hydroxy-17a-br0moprogesterone.

5. 1lfi-Hydroxy-17a-chloroprogesterone.

6. 17a-Chloro-1,9( 11)-bisdehydroprogesterone.

References Cited in the file of this patent UNITED STATES PATENTS 2,838,550 Lyttle et al June 10, 1958 

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF 17AR-9(11)-DEHYDROPROGESTERONE, 17A-R-1,9(11)-BISDEHYRDOPROGESTERONE AND 17A-R-11B-HYDROXYPROGESTERONE WHEREIN R IS A HALOGEN HAVING AN ATOMIC WEIGHT GREATER THAN 19 AND LESS THAN 126 